Functional characterization of nicotinic receptors in amacrine cells of rat retina

Autores
Gumilar, Fernanda Andrea; Politi, Luis Enrique; Bouzat, Cecilia Beatriz
Año de publicación
2008
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Amacrine cells are a heterogeneous class of interneurons that modulate the transfer of the light signals through the retina. The cholinergic system present in these neurons is involved in such modulation. However, the characterization of nicotinic cholinergic receptors (AChRs) in amacrine cells and their possible roles during the retina development remain unknown. In this study, we have identified functional AChRs in amacrine cells and determined their effects on neurite outgrowth using amacrineenriched cultures obtained from rat retinae. Amacrine cells were identified by their morphology and by immunocytochemical techniques using the monoclonal antibody HPC-1. We measured function of AChRs by electrophysiological studies and by confocal microscopy combined with the Ca2-+-sensitive fluorescent indicator Fluo-3/AM. Our results show that a7 AChRs are present in amacrine cells. The level of expression, measured by rhodamine-labelled a-bungarotoxin, increases in a time-dependent manner during culture. Maximal expression levels are achieved at days 8-9. Amacrine cells were examined with whole-cell recording techniques. Two different types of responses to ACh are observed after day 8 in culture: a slow-decay current (td = 111 +15 ms), and a fast-decay current (td = 11 = 2 ms). The latter one probably corresponds to functional a7 AChR, since this receptor shows fast desensitization and since the currents are inhibited by the specific antagonist, a-bungarotoxin. Given that a7 is highly permeable to calcium ions, we also assessed [Ca2+]i oscillations with Fluo3-AM induced by nicotinic agonists. When cells are exposed to ACh or nicotine, Ca2+ influx is detected as an increase in the fluorescence intensity. a-Bgt inhibits this signal, again indicating functional a7 AChRs. Interestingly, the presence of nicotine significant increases neuritic length of cultured amacrine cells. This outgrowth effect is abolished by co-treatment with a-B gt. In conclusion, amacrine cells have functional o7 receptors which may have a role in retinal synaptogenesis
Fil: Gumilar, Fernanda Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Politi, Luis Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Sociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiology
Bahía Blanca
Argentina
Sociedad Argentina de Investigación en Neurociéncias
Materia
NICOTINIC RECEPTORS
AMACRINE CELLS
RETINA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/242911
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/242911
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Functional characterization of nicotinic receptors in amacrine cells of rat retinaGumilar, Fernanda AndreaPoliti, Luis EnriqueBouzat, Cecilia BeatrizNICOTINIC RECEPTORSAMACRINE CELLSRETINAhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Amacrine cells are a heterogeneous class of interneurons that modulate the transfer of the light signals through the retina. The cholinergic system present in these neurons is involved in such modulation. However, the characterization of nicotinic cholinergic receptors (AChRs) in amacrine cells and their possible roles during the retina development remain unknown. In this study, we have identified functional AChRs in amacrine cells and determined their effects on neurite outgrowth using amacrineenriched cultures obtained from rat retinae. Amacrine cells were identified by their morphology and by immunocytochemical techniques using the monoclonal antibody HPC-1. We measured function of AChRs by electrophysiological studies and by confocal microscopy combined with the Ca2-+-sensitive fluorescent indicator Fluo-3/AM. Our results show that a7 AChRs are present in amacrine cells. The level of expression, measured by rhodamine-labelled a-bungarotoxin, increases in a time-dependent manner during culture. Maximal expression levels are achieved at days 8-9. Amacrine cells were examined with whole-cell recording techniques. Two different types of responses to ACh are observed after day 8 in culture: a slow-decay current (td = 111 +15 ms), and a fast-decay current (td = 11 = 2 ms). The latter one probably corresponds to functional a7 AChR, since this receptor shows fast desensitization and since the currents are inhibited by the specific antagonist, a-bungarotoxin. Given that a7 is highly permeable to calcium ions, we also assessed [Ca2+]i oscillations with Fluo3-AM induced by nicotinic agonists. When cells are exposed to ACh or nicotine, Ca2+ influx is detected as an increase in the fluorescence intensity. a-Bgt inhibits this signal, again indicating functional a7 AChRs. Interestingly, the presence of nicotine significant increases neuritic length of cultured amacrine cells. This outgrowth effect is abolished by co-treatment with a-B gt. In conclusion, amacrine cells have functional o7 receptors which may have a role in retinal synaptogenesisFil: Gumilar, Fernanda Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Politi, Luis Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaSociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiologyBahía BlancaArgentinaSociedad Argentina de Investigación en NeurociénciasSociedad Argentina de Investigación en Neurociencias2008info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectWorkshopBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/242911Functional characterization of nicotinic receptors in amacrine cells of rat retina; Sociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiology; Bahía Blanca; Argentina; 2008; 34-34CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://saneurociencias.org.arNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:43:48Zoai:ri.conicet.gov.ar:11336/242911instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:43:48.909CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Functional characterization of nicotinic receptors in amacrine cells of rat retina
title Functional characterization of nicotinic receptors in amacrine cells of rat retina
spellingShingle Functional characterization of nicotinic receptors in amacrine cells of rat retina
Gumilar, Fernanda Andrea
NICOTINIC RECEPTORS
AMACRINE CELLS
RETINA
title_short Functional characterization of nicotinic receptors in amacrine cells of rat retina
title_full Functional characterization of nicotinic receptors in amacrine cells of rat retina
title_fullStr Functional characterization of nicotinic receptors in amacrine cells of rat retina
title_full_unstemmed Functional characterization of nicotinic receptors in amacrine cells of rat retina
title_sort Functional characterization of nicotinic receptors in amacrine cells of rat retina
dc.creator.none.fl_str_mv Gumilar, Fernanda Andrea
Politi, Luis Enrique
Bouzat, Cecilia Beatriz
author Gumilar, Fernanda Andrea
author_facet Gumilar, Fernanda Andrea
Politi, Luis Enrique
Bouzat, Cecilia Beatriz
author_role author
author2 Politi, Luis Enrique
Bouzat, Cecilia Beatriz
author2_role author
author
dc.subject.none.fl_str_mv NICOTINIC RECEPTORS
AMACRINE CELLS
RETINA
topic NICOTINIC RECEPTORS
AMACRINE CELLS
RETINA
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Amacrine cells are a heterogeneous class of interneurons that modulate the transfer of the light signals through the retina. The cholinergic system present in these neurons is involved in such modulation. However, the characterization of nicotinic cholinergic receptors (AChRs) in amacrine cells and their possible roles during the retina development remain unknown. In this study, we have identified functional AChRs in amacrine cells and determined their effects on neurite outgrowth using amacrineenriched cultures obtained from rat retinae. Amacrine cells were identified by their morphology and by immunocytochemical techniques using the monoclonal antibody HPC-1. We measured function of AChRs by electrophysiological studies and by confocal microscopy combined with the Ca2-+-sensitive fluorescent indicator Fluo-3/AM. Our results show that a7 AChRs are present in amacrine cells. The level of expression, measured by rhodamine-labelled a-bungarotoxin, increases in a time-dependent manner during culture. Maximal expression levels are achieved at days 8-9. Amacrine cells were examined with whole-cell recording techniques. Two different types of responses to ACh are observed after day 8 in culture: a slow-decay current (td = 111 +15 ms), and a fast-decay current (td = 11 = 2 ms). The latter one probably corresponds to functional a7 AChR, since this receptor shows fast desensitization and since the currents are inhibited by the specific antagonist, a-bungarotoxin. Given that a7 is highly permeable to calcium ions, we also assessed [Ca2+]i oscillations with Fluo3-AM induced by nicotinic agonists. When cells are exposed to ACh or nicotine, Ca2+ influx is detected as an increase in the fluorescence intensity. a-Bgt inhibits this signal, again indicating functional a7 AChRs. Interestingly, the presence of nicotine significant increases neuritic length of cultured amacrine cells. This outgrowth effect is abolished by co-treatment with a-B gt. In conclusion, amacrine cells have functional o7 receptors which may have a role in retinal synaptogenesis
Fil: Gumilar, Fernanda Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Politi, Luis Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Sociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiology
Bahía Blanca
Argentina
Sociedad Argentina de Investigación en Neurociéncias
description Amacrine cells are a heterogeneous class of interneurons that modulate the transfer of the light signals through the retina. The cholinergic system present in these neurons is involved in such modulation. However, the characterization of nicotinic cholinergic receptors (AChRs) in amacrine cells and their possible roles during the retina development remain unknown. In this study, we have identified functional AChRs in amacrine cells and determined their effects on neurite outgrowth using amacrineenriched cultures obtained from rat retinae. Amacrine cells were identified by their morphology and by immunocytochemical techniques using the monoclonal antibody HPC-1. We measured function of AChRs by electrophysiological studies and by confocal microscopy combined with the Ca2-+-sensitive fluorescent indicator Fluo-3/AM. Our results show that a7 AChRs are present in amacrine cells. The level of expression, measured by rhodamine-labelled a-bungarotoxin, increases in a time-dependent manner during culture. Maximal expression levels are achieved at days 8-9. Amacrine cells were examined with whole-cell recording techniques. Two different types of responses to ACh are observed after day 8 in culture: a slow-decay current (td = 111 +15 ms), and a fast-decay current (td = 11 = 2 ms). The latter one probably corresponds to functional a7 AChR, since this receptor shows fast desensitization and since the currents are inhibited by the specific antagonist, a-bungarotoxin. Given that a7 is highly permeable to calcium ions, we also assessed [Ca2+]i oscillations with Fluo3-AM induced by nicotinic agonists. When cells are exposed to ACh or nicotine, Ca2+ influx is detected as an increase in the fluorescence intensity. a-Bgt inhibits this signal, again indicating functional a7 AChRs. Interestingly, the presence of nicotine significant increases neuritic length of cultured amacrine cells. This outgrowth effect is abolished by co-treatment with a-B gt. In conclusion, amacrine cells have functional o7 receptors which may have a role in retinal synaptogenesis
publishDate 2008
dc.date.none.fl_str_mv 2008
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Workshop
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/242911
Functional characterization of nicotinic receptors in amacrine cells of rat retina; Sociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiology; Bahía Blanca; Argentina; 2008; 34-34
CONICET Digital
CONICET
url http://hdl.handle.net/11336/242911
identifier_str_mv Functional characterization of nicotinic receptors in amacrine cells of rat retina; Sociedad Argentina de Investigación en Neurociencias. Workshop Neuronal Communication: From structure to physiology; Bahía Blanca; Argentina; 2008; 34-34
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://saneurociencias.org.ar
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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